A time-course study of the effect of sulphur on glucosinolates in oilseed rape (Brassica napus) from the vegetative stage to maturity

Samples consisting of the whole above-ground part of the oilseed rape plant (Brassica napus L), were taken every 2 weeks from before stem extension to maturity. The plants were separated into vegetative tissue, floral tissue, pods and seeds (when these components appeared), and the individual glucosinolates present were quantified by high performance liquid chromatography. A high glucosinolate variety (Rafal) and a low glucosinolate variety (Cobra) were compared. The effect of sulphur (32 kg ha^?1 elemental sulphur applied at the beginning of stem extension) on the various parts of the plant was investigated. Total glucosinolate concentration in all plant parts was higher in Rafal than in Cobra. Glucosinolate concentration in the vegetation declined over time, most noticeably when the flowers and seeds were produced, and glucosinolate concentration of the pods also fell as that of the seeds rose. It is suggested that there may have been some redistribution of glucosinolates or glucosinolate precursors within the plant as maturity approached. Changes occurred in the proportions that individual glucosinolates contributed to the total glucosinolate content, and this may be relevant to plant/pathogen relationships. Sulphur application increased the glucosinolate concentration of the vegetative tissue by mid April and also increased the glucosinolate concentration of the flowers. It is suggested that this could affect the plant's resistance to disease.     

Interactions of Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes plants cultivated in a gnotobiotic system

The growth and persistence of Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes on a diverse range of plant types over extended cultivation periods was studied. When introduced on the seed of carrot, cress, lettuce, radish, spinach and tomato all the pathogens became rapidly established shortly after germination, attaining cell densities of the order of 5.5-6.5 log cfu/g. In general, Es. coli O157:H7 and L. monocytogenes became established and persisted at significantly higher levels on seedlings (9 days post-germination) than Salmonella. Es. coli O157:H7 became internalized in cress, lettuce, radish and spinach seedlings but was not recovered within the tissues of mature plants. Internalization of Salmonella was also observed in lettuce and radish but not cress or spinach seedlings. In contrast, L. monocytogenes did not internalize within seedlings but did persist on the surface of plants throughout the cultivation period. Co-inoculation of isolates recovered from the rhizosphere of plants did not significantly affect the numbers or persistence of human pathogens. The only exception was with Enterobacter cloacae, which reduced Es. coli O157:H7 Ph1 and L. monocytogenes levels by ca. 1 log cfu/g on lettuce. With the bioluminescent phenotype of Es. coli O157:H7 Ph1, it was demonstrated that the human pathogen became established on the roots of growing plants. Scanning electron micrographs of root seedlings suggested that Es. coli O157:H7 Ph1 preferentially colonized the root junctions of seedlings. It is proposed that such colonization sites enhanced the persistence of Es. coli O157:H7 on plants and facilitated internalization within developing seedlings. The results suggest that the risk associated with internalized human pathogens in salad vegetables at harvest is low. Nevertheless, the introduction of human pathogens at an early stage of plant development could enhance their persistence in the rhizosphere. The implications of the study with regards to on-farm food safety initiatives are discussed.     

Internalization of Escherichia coli O157:H7 following biological and mechanical disruption of growing spinach plants

The internalization and persistence of a bioluminescent Escherichia coli O157:H7 Ph1 was investigated in growing spinach plants that had been either biologically or mechanically damaged. In control (undamaged) plants cultivated in soil microcosms inoculated with E. coli O157:H7 Phl, the bacterium was recovered from surface-sterilized root tissue but not from leaves. Mechanical disruption of the seminal root and root hairs of the plants did not result in the internalization of the pathogen into the aerial leaf tissue. When imprints of the root tissue were made on plates of tryptic soy agar plus ampicillin, no colonies of E. coli O157:H7 were observed around damaged tissue. The roots of growing plants were exposed to the northern root-knot nematode, Meloidogyne hapla, in the presence of E. coli O157:H7. Although this treatment caused knot formation on the roots, it did not enhance the internalization of the bacterium into the plant vascular system. Coinoculation of intact leaves with E. coli O157:H7 and the phytopathogen Pseudomonas syringae DC3000 resulted in localized necrosis, but the persistence of the human pathogen was not affected. The mechanical disruption of roots does not result in the internalization of E. coli O157:H7 into the aerial tissue of spinach, and there does not appear to be any effect of P. syringae in terms of enhancing the persistence of E. coli O157:H7 in spinach leaves.     

Sensitivity to change in laminar burning velocity and Markstein length resulting from variable hydrogen fraction in blast furnace gas for changing ambient conditions

The sensitivity to changes in fuel characteristics has been investigated for combustion of Blast Furnace Gas resulting from small volumetric increases in H₂ concentration. A nonlinear methodology has been employed to quantify unstretched flame speeds and the effect of flame stretch from outwardly propagating spherical flames. Following benchmarking work with CH₄, results were obtained under ambient conditions of 303 K and 0.1 MPa, with small absolute change in hydrogen concentration shown to at least triple the laminar burning velocity for all tested mixtures. Fuel composition and equivalence ratio were shown to independently influence mixture diffusivity and Lewis number, quantified by change in the obtained values of Markstein length. Temperature and pressure were increased to respective values of 393 K and 0.2 MPa to investigate influence of ambient conditions, with a power law correlation presented. Finally the performance of several published chemical reaction mechanisms has been evaluated through comparison of 1-D flame models.     

Chain length of the polylysine in receptor-targeted gene transfer complexes affects duration of reporter gene expression both in vitro and in vivo

Complexes composed of peptide ligand for the serpin enzyme complex receptor covalently coupled to poly-L-lysine condensed by charge interaction with plasmid DNA direct gene transfer into receptor bearing cells. We compared intensity and duration of reporter gene expression in vitro and in vivo from serpin-enzyme receptor-directed gene transfer complexes prepared with poly-L-lysine of different chain lengths. When substituted with linker and ligand to comparable extents, DNA complexes containing short chain poly-L-lysine were larger and gave higher peak expression but significantly shorter duration of expression than those containing long chain poly-L-lysine. Both peak expression and duration of expression exceeded that observed with Lipofectin. Neither naked DNA nor DNA complexed with unsubstituted polylysine was effective in gene transfer. For in vivo experiments, complexes containing optimal ligand and degree of substitution (based on in vitro data, peptide C105Y, 11 ligands/plasmid DNA molecule) were prepared with either short chain or long chain polylysine and a beta-galactosidase expression plasmid. Following injection into the tail veins of mice, longer chain complexes gave significantly higher expression of reporter gene in lung and spleen that lasted for a significantly longer period of time than the shorter chain complexes. The short chain poly-L-lysine-DNA complexes were larger in diameter, as assessed by electron microscopy or atomic force microscopy, and gave less protection against DNase digestion in vitro than longer chain complexes. Thus, for gene transfer complexes directed at the serpin enzyme complex receptor, longer chain poly-L-lysine gave a much longer duration of expression both in vitro and in vivo. We speculate that this may be due to protection against degradation afforded the plasmid DNA by the tighter compaction produced by long chain poly-L-lysine.     

GS32, a novel Golgi SNARE of 32 kDa, interacts preferentially with syntaxin 6

Syntaxin 1, synaptobrevins or vesicle-associated membrane proteins, and the synaptosome-associated protein of 25 kDa (SNAP-25) are key molecules involved in the docking and fusion of synaptic vesicles with the presynaptic membrane. We report here the molecular, cell biological, and biochemical characterization of a 32-kDa protein homologous to both SNAP-25 (20% amino acid sequence identity) and the recently identified SNAP-23 (19% amino acid sequence identity). Northern blot analysis shows that the mRNA for this protein is widely expressed. Polyclonal antibodies against this protein detect a 32-kDa protein present in both cytosol and membrane fractions. The membrane-bound form of this protein is revealed to be primarily localized to the Golgi apparatus by indirect immunofluorescence microscopy, a finding that is further established by electron microscopy immunogold labeling showing that this protein is present in tubular-vesicular structures of the Golgi apparatus. Biochemical characterizations establish that this protein behaves like a SNAP receptor and is thus named Golgi SNARE of 32 kDa (GS32). GS32 in the Golgi extract is preferentially retained by the immobilized GST-syntaxin 6 fusion protein. The coimmunoprecipitation of syntaxin 6 but not syntaxin 5 or GS28 from the Golgi extract by antibodies against GS32 further sustains the preferential interaction of GS32 with Golgi syntaxin 6.     

An unconventional role for cytoplasmic disulfide bonds in vaccinia virus proteins

Previous data have shown that reducing agents disrupt the structure of vaccinia virus (vv). Here, we have analyzed the disulfide bonding of vv proteins in detail. In vv-infected cells cytoplasmically synthesized vv core proteins became disulfide bonded in the newly assembled intracellular mature viruses (IMVs). vv membrane proteins also assembled disulfide bonds, but independent of IMV formation and to a large extent on their cytoplasmic domains. If disulfide bonding was prevented, virus assembly was only partially impaired as shown by electron microscopy as well as a biochemical assay of IMV formation. Under these conditions, however, the membranes around the isolated particles appeared less stable and detached from the underlying core. During the viral infection process the membrane proteins remained disulfide bonded, whereas the core proteins were reduced, concomitant with delivery of the cores into the cytoplasm. Our data show that vv has evolved an unique system for the assembly of cytoplasmic disulfide bonds that are localized both on the exterior and interior parts of the IMV.     

Competing strategies in categorization: Expediency and resistance to knowledge restructuring.

The authors investigated people's ability to restructure their knowledge when additional information about a categorization task is revealed. In 2 experiments, people first learned to rely on a fairly accurate (but imperfect) predictor. At various points in training, a complex relationship between 2 other predictors was revealed in a schematic diagram that could support perfect performance. In Experiment 1, people adopted the complex strategy when it was revealed at the outset but were unable to restructure their knowledge after the expedient predictor had been learned. In Experiment 2, expedient knowledge persisted even with an adaptive display. The persistence of expedient knowledge is explained by associative blocking of potential alternative cues. A 3rd experiment analyzed the strategies people use with and without the diagram. The study confirmed that the diagram, when presented at the outset, significantly alters people's approach to the task. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Technical information

The centre of glass thermal performance of evacuated glazing is better than the performance of argon‐filled atmospheric pressure double glazings. Mitigation, or elimination of evacuated glazing edge effects will bring total window thermal performance on par, or better than, competing ‘superwindow’ options; thus increasing the commercial attractiveness of these glazings. The results of the computer‐based thermal modelling at the Universities of Sydney and Ulster compare well with each other and with the infrared thermography measurements at Lawrence Berkeley.